Analysis of eco-environmental water requirements (EEWRs) and water resource allocation strategies for arid, inland river basins can provide the theoretical basis for sustainable water utilization and management. In this paper, an optimal water resource allocation strategy is proposed for Yarkand River Basin in Xinjiang, China, on the basis of a comprehensive analysis of runoff data collected between 1970 and 2016, three ecological environmental protection goals, basic eco-environmental water requirement (BEEWR) aimed at sustaining aquatic ecosystems within the river, and target eco-environmental water requirements (TEEWR) aimed at protecting various types of riparian vegetation along the river. The results showed that: (1) after the runoff in Kaqun reach subtracting the BEEWR, the annual average river loss (recharge), and the amount of water diversion for irrigation (51.43 × 10⁸ m³) from flows along the Kaqun reach, the remaining water volume during wet years was able to meet all three TEEWRs; (2) during moderately wet years, the remaining water was capable of meeting the second and third TEEWRs; and (3) during dry and extremely dry years, there was little or no residual water available to meet TEEWRs. The proposed optimal water resource allocation strategy, based on the above findings, states that the water diversion requirement for irrigation and domestic use allocated from the total amount of runoff should not exceed the National Water Policy (Three Red Lines) standard first. Then, the BEEWR allocated from the runoff should be met second, and the annual average river loss, third. Depending on the amount of remaining water, the second and third TEEWRs can be fulfilled during wet years, but during moderately wet years, only the third TEEWR can be met. During dry and extremely dry years, only the BEEWR of the river can be met and only during the flood season.